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Literature summary for 2.4.2.28 extracted from

  • Cacciapuoti, G.; Servillo, L.; Moretti, M.A.; Porcelli, M.
    Conformational changes and stabilization induced by phosphate binding to 5'-methylthioadenosine phosphorylase from the thermophilic archaeon Sulfolobus solfataricus (2001), Extremophiles, 5, 295-302.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
overexpression in Escherichia coli strain RB791, amino acid determination, incorrect positioning of disulfide bonds, the recombinant enzyme is less thermostable and thermophilic than the native enzyme Saccharolobus solfataricus

Crystallization (Commentary)

Crystallization (Comment) Organism
from recombinant enzyme, hanging drop-vapour diffusion method, protein solution, 7-10 mg/ml, 18°C, reservoir solution for native crystals: Tris-HCl 10 mM, pH 7.4, 28-30% dioxane, 12% 2-methyl-2,4-pentanediol, 0.12 M MgCl2, 0,04 M NaCl, for crystals of enzyme complexed with substrates or sulfate and phosphate ions, substrates are added and NaCl is exchanged for MgSO4 or NH4Cl and KH2PO4, respectively, X-ray structure determination and analysis Saccharolobus solfataricus

General Stability

General Stability Organism
phosphate, and less efficiently also arsenate and sulfate, stabilize the recombinant enzyme against inactiviation by temperature, SDS, urea, and proteolytic enzymes Saccharolobus solfataricus
recombinant enzyme, 90°C in 2% SDS, 30 min, loss of 60% activity Saccharolobus solfataricus
recombinant enzyme, 90°C in 8 M urea, 30 min, loss of 70% activity Saccharolobus solfataricus

Inhibitors

Inhibitors Comment Organism Structure
proteinase K recombinant enzyme, 10% remaining activity after 4 h at 37°C, phosphate protects Saccharolobus solfataricus
Subtilisin recombinant enzyme, 24% remaining activity after 4 h at 37°C, phosphate protects Saccharolobus solfataricus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
27000
-
6 * 27000, recombinant enzyme, crystal structure analysis Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant from Escherichia coli Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5'-methylthioadenosine + phosphate binding of phosphate and 5-methylthioribose 1-phosphate to the enzyme induces a conformational transition that stabilizes the folded structure of the enzyme Saccharolobus solfataricus adenine + 5-methylthio-D-ribose 1-phosphate
-
?
5'-methylthioadenosine + phosphate 5'-methylthioadenosine and adenine form ternary complexes with the enzyme only in presence of phosphate and methylthioribose 1-phosphate, respectively Saccharolobus solfataricus adenine + 5-methylthio-D-ribose 1-phosphate
-
?

Subunits

Subunits Comment Organism
hexamer 6 * 27000, recombinant enzyme, crystal structure analysis Saccharolobus solfataricus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
100
-
recombinant enzyme, 1 h, 95% remainig activity in presence of 100 mM phosphate Saccharolobus solfataricus
110
-
recombinant enzyme, 10 min, 50% remaining activity in absence and 90% remaining activity in presence of 100 mM phosphate Saccharolobus solfataricus
120
-
recombinant enzyme, 10 min, no activity in absence and 50% remaining activity in presence of 100 mM phosphate Saccharolobus solfataricus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Saccharolobus solfataricus